Abstract：
:Spatiotemporal activity patterns in three-dimensionally organized cellular networks are fundamental to the function of the nervous system. Despite advances in functional imaging of cell populations, a method to resolve local network activity in three dimensions has been lacking. Here we introduce a three-dimensional (3D) line-scan technology for two-photon microscopy that permits fast fluorescence measurements from several hundred cells distributed in 3D space. We combined sinusoidal vibration of the microscope objective at 10 Hz with 'smart' movements of galvanometric x-y scanners to repeatedly scan the laser focus along a closed 3D trajectory. More than 90% of cell somata were sampled by the scan line within volumes of 250 microm side length. Using bulk-loading of calcium indicator, we applied this method to reveal spatiotemporal activity patterns in neuronal and astrocytic networks in the rat neocortex in vivo. Two-photon population imaging using 3D scanning opens the field for comprehensive studies of local network dynamics in intact tissue.
journal_name
Nat Methodsjournal_title
Nature methodsauthors
Göbel W,Kampa BM,Helmchen Fdoi
10.1038/nmeth989subject
Has Abstractpub_date
2007-01-01 00:00:00pages
73-9issue
1eissn
1548-7091issn
1548-7105pii
nmeth989journal_volume
4pub_type
杂志文章相关文献
NATURE METHODS文献大全abstract：:High spatial and temporal resolution of conditional gene expression is typically difficult to achieve in whole tissues or organisms. We synthesized two reversibly inhibited, photoactivatable ('caged') doxycycline derivatives with different membrane permeabilities for precise spatial and temporal light-controlled activ...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1340
更新日期：2009-07-01 00:00:00
abstract：:Stable in vivo mapping and modulation of the same neurons and brain circuits over extended periods is critical to both neuroscience and medicine. Current electrical implants offer single-neuron spatiotemporal resolution but are limited by such factors as relative shear motion and chronic immune responses during long-t...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3969
更新日期：2016-10-01 00:00:00
abstract：:Electrically excitable cells are important in the normal functioning and in the pathophysiology of many biological processes. These cells are typically embedded in dense, heterogeneous tissues, rendering them difficult to target selectively with conventional electrical stimulation methods. The algal protein Channelrho...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth936
更新日期：2006-10-01 00:00:00
abstract：:Single-molecule localization microscopy is a powerful tool for visualizing subcellular structures, interactions and protein functions in biological research. However, inhomogeneous refractive indices inside cells and tissues distort the fluorescent signal emitted from single-molecule probes, which rapidly degrades res...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-020-0816-x
更新日期：2020-05-01 00:00:00
abstract：:We developed a method that allows release of intact membrane protein complexes from amphipols, bicelles and nanodiscs in the gas phase for observation by mass spectrometry (MS). Current methods involve release of membrane protein complexes from detergent micelles, which reveals subunit composition and lipid binding. W...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.2691
更新日期：2013-12-01 00:00:00
abstract：:We present general means to greatly increase the sensitivity of antibody-based assays. Augmentation relies on a 'tadpole' protein-DNA chimera whose protein moiety binds most classes of mammalian antibodies but not avian immunoglobulin Y (IgY). We used this tadpole in affinity capture assays followed by real-time PCR t...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth1127
更新日期：2007-12-01 00:00:00
abstract：:Infrared fluorescent proteins (IFPs) provide an additional color to GFP and its homologs in protein labeling. Drawing on structural analysis of the dimer interface, we identified a bacteriophytochrome in the sequence database that is monomeric in truncated form and engineered it into a naturally monomeric IFP (mIFP). ...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3447
更新日期：2015-08-01 00:00:00
abstract：:We introduce field synthesis, a theorem and method that can be used to synthesize any scanned or dithered light sheet, including those used in lattice light-sheet microscopy (LLSM), from an incoherent superposition of one-dimensional intensity distributions. Compared to LLSM, this user-friendly and modular approach of...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-019-0327-9
更新日期：2019-03-01 00:00:00
abstract：:RNA-guided Cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have dramatically transformed our ability to edit the genomes of diverse organisms. We believe tools and techniques based on Cas9, a single unifying factor capable of colocalizing RNA, DNA and protein,...
journal_title：Nature methods
pub_type： 杂志文章,评审
doi：10.1038/nmeth.2649
更新日期：2013-10-01 00:00:00
abstract：:Transcriptional enhancers are a primary mechanism by which tissue-specific gene expression is achieved. Despite the importance of these regulatory elements in development, responses to environmental stresses and disease, testing enhancer activity in animals remains tedious, with a minority of enhancers having been cha...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.2558
更新日期：2013-08-01 00:00:00
abstract：:We have developed hydrogel-based virtual microfluidics as a simple and robust alternative to complex engineered microfluidic systems for the compartmentalization of nucleic acid amplification reactions. We applied in-gel digital multiple displacement amplification (dMDA) to purified DNA templates, cultured bacterial c...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3955
更新日期：2016-09-01 00:00:00
abstract：:Targeted quantification of DNA methylation allows for interrogation of the most informative loci across many samples quickly and cost-effectively. Here we report improved bisulfite padlock probes (BSPPs) with a design algorithm to generate efficient padlock probes, a library-free protocol that dramatically reduces sam...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1871
更新日期：2012-02-05 00:00:00
abstract：:The detection of somatic mutations from cancer genome sequences is key to understanding the genetic basis of disease progression, patient survival and response to therapy. Benchmarking is needed for tool assessment and improvement but is complicated by a lack of gold standards, by extensive resource requirements and b...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3407
更新日期：2015-07-01 00:00:00
abstract：:We present functional ultrasound (fUS), a method for imaging transient changes in blood volume in the whole brain at better spatiotemporal resolution than with other functional brain imaging modalities. fUS uses plane-wave illumination at high frame rate and can measure blood volumes in smaller vessels than previous u...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1641
更新日期：2011-07-03 00:00:00
abstract：: ...
journal_title：Nature methods
pub_type： 评论,杂志文章
doi：10.1038/s41592-018-0248-z
更新日期：2018-12-01 00:00:00
abstract：:Our Patterning on Topography (PoT) printing technique enables fibronectin, laminin and other proteins to be applied to biomaterial surfaces in complex geometries that are inaccessible using traditional soft lithography techniques. Engineering combinatorial surfaces that integrate topographical and biochemical micropat...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3210
更新日期：2015-02-01 00:00:00
abstract：:We describe Trans-ABySS, a de novo short-read transcriptome assembly and analysis pipeline that addresses variation in local read densities by assembling read substrings with varying stringencies and then merging the resulting contigs before analysis. Analyzing 7.4 gigabases of 50-base-pair paired-end Illumina reads f...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1517
更新日期：2010-11-01 00:00:00
abstract：:Microarrays have been widely used for the analysis of gene expression, but the issue of reproducibility across platforms has yet to be fully resolved. To address this apparent problem, we compared gene expression between two microarray platforms: the short oligonucleotide Affymetrix Mouse Genome 430 2.0 GeneChip and a...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth757
更新日期：2005-05-01 00:00:00
abstract：:Circular RNAs (circRNAs) produced from back-spliced exons are widely expressed, but individual circRNA functions remain poorly understood owing to the lack of adequate methods for distinguishing circRNAs from cognate messenger RNAs with overlapping exons. Here, we report that CRISPR-RfxCas13d can effectively discrimin...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-020-01011-4
更新日期：2021-01-01 00:00:00
abstract：:Cortical information processing relies on synaptic interactions between diverse classes of neurons with distinct electrophysiological and connection properties. Uncovering the operational principles of these elaborate circuits requires the probing of electrical activity from selected populations of defined neurons. He...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1479
更新日期：2010-08-01 00:00:00
abstract：: ...
journal_title：Nature methods
pub_type： 评论,杂志文章
doi：10.1038/s41592-019-0343-9
更新日期：2019-03-01 00:00:00
abstract：:Systems biology conceptualizes biological systems as dynamic networks of interacting elements, whereby functionally important properties are thought to emerge from the structure of such networks. Owing to the ubiquitous role of complexes of interacting proteins in biological systems, their subunit composition and temp...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.1390
更新日期：2009-11-01 00:00:00
abstract：:Extracellular vesicles (EVs) are secreted nanosized particles with many biological functions and pathological associations. The inability to image EVs in fixed tissues has been a major limitation to understanding their role in healthy and diseased tissue microenvironments. Here, we show that crosslinking mammalian tis...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-019-0623-4
更新日期：2019-12-01 00:00:00
abstract：:Numerous drugs and endogenous ligands bind to cell surface receptors leading to modulation of downstream signaling cascades and frequently to adaptation of the plasma membrane proteome. In-depth analysis of dynamic processes at the cell surface is challenging due to biochemical properties and low abundances of plasma ...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/s41592-020-01022-1
更新日期：2021-01-01 00:00:00
abstract：:We introduce PyClone, a statistical model for inference of clonal population structures in cancers. PyClone is a Bayesian clustering method for grouping sets of deeply sequenced somatic mutations into putative clonal clusters while estimating their cellular prevalences and accounting for allelic imbalances introduced ...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.2883
更新日期：2014-04-01 00:00:00
abstract：:The reconstruction of neuronal populations, a key step in understanding neural circuits, remains a challenge in the presence of densely packed neurites. Here we achieved automatic reconstruction of neuronal populations by partially mimicking human strategies to separate individual neurons. For populations not resolvab...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.3662
更新日期：2016-01-01 00:00:00
abstract：:Understanding how cells maintain genome integrity when challenged with DNA double-strand breaks (DSBs) is of major importance, particularly since the discovery of multiple links of DSBs with genome instability and cancer-predisposition disorders. Ionizing radiation is the agent of choice to produce DSBs in cells; howe...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.f.206
更新日期：2008-03-01 00:00:00
abstract：:The ability to selectively interfere with post-translationally modified proteins would have many biological and therapeutic applications. However, post-translational modifications cannot be selectively targeted by nucleic-acid-based interference approaches. Here we describe post-translational intracellular silencing a...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.4144
更新日期：2017-03-01 00:00:00
abstract：:We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy with GFP-labeled samples. The approximately 70 nm lateral resolution attained in this study is demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell. Our results mark the...
journal_title：Nature methods
pub_type： 杂志文章,评审
doi：10.1038/nmeth922
更新日期：2006-09-01 00:00:00
abstract：:Automated annotation of protein function is challenging. As the number of sequenced genomes rapidly grows, the overwhelming majority of protein products can only be annotated computationally. If computational predictions are to be relied upon, it is crucial that the accuracy of these methods be high. Here we report th...
journal_title：Nature methods
pub_type： 杂志文章
doi：10.1038/nmeth.2340
更新日期：2013-03-01 00:00:00
